LOCUS       sequence-232275-        6144 bp    DNA     circular SYN 14-APR-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6 tag, cleavable with 3C,
            and N-terminal mRuby3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  AUTHORS   Lemaitre RP, Bogdanova A, Borgonovo B, Woodruff JB, Drechsel DN
  TITLE     FlexiBAC: a versatile, open-source baculovirus vector system for 
            protein expression, secretion, and proteolytic processing.
  JOURNAL   BMC Biotechnol. 2019 Mar 29;19(1):20. doi: 
            10.1186/s12896-019-0512-z.
  PUBMED    30925874
REFERENCE   2  (bases 1 to 6144)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol."; date: "2019-03-29"; volume: "19(1):20. doi"; pages: " 
            10.1186/s12896-019-0512-z"
COMMENT     Sequence Label: pOCC291
FEATURES             Location/Qualifiers
     source          1..6144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      134..143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             152..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             182..205
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             206..916
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     CDS             1055..1360
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1239..1258
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1533..1667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1570..1589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1624..1643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1740..2112
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2464..2481)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2574..3162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2654..2673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3333..4124,4125..4193))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3956..3975
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4194..4298)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4599..4621
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4613..4630
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4614..4630
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4641..4660
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4641..4659
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5270..5394
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5432..6037)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcgccacca tgggcagcag ccatcaccat caccatcaca gcagcggcag
      181 actggaagtt ctgttccagg ggccgatggt gtctaagggc gaagagctga tcaaggaaaa
      241 tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc caccaattca aatgcacagg
      301 tgaaggagaa ggcagaccgt acgagggaac tcaaaccatg aggatcaaag tcatcgaggg
      361 aggacccctg ccatttgcct ttgacattct tgccacgtcg ttcatgtatg gcagccgtac
      421 ttttatcaag tacccggccg acatccctga tttctttaaa cagtcctttc ctgagggttt
      481 tacttgggaa agagttacga gatacgaaga tggtggagtc gtcaccgtca cgcaggacac
      541 cagccttgag gatggcgagc tcgtctacaa cgtcaaggtc agaggggtaa actttccctc
      601 caatggtccc gtgatgcaga agaagaccaa gggttgggag cctaatacag agatgatgta
      661 tccagcagat ggtggtctga gaggatacac tgacatcgca ctgaaagttg atggtggtgg
      721 ccatctgcac tgcaacttcg tgacaactta caggtcaaaa aagaccgtcg ggaacatcaa
      781 gatgcccggt gtccatgccg ttgatcaccg cctggaaagg atcgaggaga gtgacaatga
      841 aacctacgta gtgcaacgcg aagtggcagt tgccaaatac agcaaccttg gtggtggcat
      901 ggacgagctg tacaagggat ccgctggctc cgctgctggt tctggcgcgg ccgcaagcgc
      961 tatggccaca acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact
     1021 gtctgcttac ataaacagta atacaagggg tgttatgcag tttaaggttt acacctataa
     1081 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     1141 gcgacggatg gtgatccccc tggccagtgc ccgtctgctg tcagataaag tctcccgtga
     1201 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     1261 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     1321 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     1381 cagccagtgg cgcgccgtaa tagaagcttg tcgagaagta ctagaggatc ataatcagcc
     1441 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1501 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1561 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1621 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg
     1681 agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt aattaataaa
     1741 acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta
     1801 cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg
     1861 ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct
     1921 caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga
     1981 tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca
     2041 gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt
     2101 tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg
     2161 acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta gctttattag
     2221 gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg
     2281 ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag
     2341 ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt cgggaaacct
     2401 gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc
     2461 ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga
     2521 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2581 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2641 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2701 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2761 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2821 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2881 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2941 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3001 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3061 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3121 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3181 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3241 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3301 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3361 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3421 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
     3481 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3541 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3601 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3661 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3721 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3781 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3841 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3901 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3961 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4021 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4081 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4141 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4201 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4261 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4321 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4381 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4441 agatagggtt gagtggccgc tacagggcgc tcccattcgc cattcaggct gcgcaactgt
     4501 tgggaagggc gtttcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4561 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     4621 gacggccagt gagcgcgacg taatacgact cactataggg cgaattggcg gaaggccgtc
     4681 aaggccgtga cgttaaaact attaagccat ccaatcgacc gttagtcgaa tcaggaccgc
     4741 tggtgcgaga agccgcgaag tatggcgaat gcatcgtata acgtgtggag tccgctcatt
     4801 agagcgtcat gtttagacaa gaaagctaca tatttaattg atcccgatga ttttattgat
     4861 aaattgaccc taactccata cacggtattc tacaatggcg gggttttggt caaaatttcc
     4921 ggactgcgat tgtacatgct gttaacggct ccgcccacta ttaatgaaat taaaaattcc
     4981 aattttaaaa aacgcagcaa gagaaacatt tgtatgaaag aatgcgtaga aggaaagaaa
     5041 aatgtcgtgg acatgctgaa caacaagatt aatatgcctc cgtgtataaa aaaaatattg
     5101 aacgatttga aagaaaacaa tgtaccgcgc ggcggtatgt acaggaagag gtttatacta
     5161 aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag
     5221 gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta
     5281 atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac tgtggacaag
     5341 ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta ttgaataata
     5401 aaacaattat aaatgctaaa tttgtttttt attaacgata caaaccaaac gcaacaagaa
     5461 catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg taatatttaa
     5521 aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt tcacataaac
     5581 tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc tcctcataaa
     5641 aattaacata gttattatcg tatccatata tgtatctatc gtatagagta aattttttgt
     5701 tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc atagtttttc
     5761 tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct ttaattatta
     5821 aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg ccggcatagt
     5881 acgcagcttc ttctagttca attacaccat tttttagcag caccggatta acataacttt
     5941 ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct atactattgt
     6001 ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc acaaactaat
     6061 atcacaagct gccaccattt tgttcgccca ggactctagc tatagttcta gtggttggct
     6121 acgtatactc cggaatatta atag
//
